Anti ampkα

All chemicals were purchased from Sigma–Aldrich (St Louis, MO, USA) unless otherwise noted. Compound C (CC) also named Dorsomorphin: 6-[4-(2-Piperidin-1-yl-ethoxy)-phenyl]-3-pyridin-4-yl-pyrazolo[1,5-a]pyrimidine, AICAR: 5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside (AICAR), and Metformin (MET): 1,1-dimethylbiguanide hydrochloride were obtained from Calbiochem (Billerica, MA). A stock solution of CC was prepared in dimethylsulphoxide (DMSO) and stock solutions of MET and AICAR were prepared in deionized water. Complete mini EDTA-free, protease inhibitor cocktail tablets were from Roche diagnostics (Mannheim, Germany). Tris/glycine buffer (10X), Tris/glycine/SDS buffer (10X), and Precision Plus Protein All Blue Standards (Catalog #161–0373) were obtained from Bio-Rad (Hercules, CA) and anti-AMPKα from Millipore (Billerica, MA), anti-phospho-Thr¹⁷²-AMPKα and anti-rabbit IgG (H+L) (DyLight 680 Conjugate) antibodies from Cell Signaling technology, Inc (Danvers, MA). SYBR-14/PI (LIVE/DEAD sperm viability kit) and JC1 were purchased from Molecular Probes (Saint Aubin, France). The LPO-586 kit was from Oxis Research (Burlingame, CA, US).

Nitrocellulose membrane, polyacrylamide solution and protein standards were purchased from Bio-Rad Laboratories (Hercules, CA, USA). Antibodies against phospho-extracellular signal-regulated protein kinase p-ERK [T202/Y204], phospho-AMP-activated protein kinase p-AMPK [T172], phospho-p38 mitogen-activated protein (MAP) kinase p-p38 [T180/Y182], phospho-ribosomal protein S6 p-S6 [S235/S236] and S6 were obtained from Cell Signaling Technology (Beverly, MA, USA). Anti-p70S6 kinase or 70 kDa ribosomal protein S6 kinase S6K1 [T389], anti-p38α and anti-ERKα antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA), anti-vinculin from Sigma Chemical Company (St Louis, MO, USA) and anti-AMPKα from Millipore (Paris, France). Alexa Fluor secondary antibodies were purchased from Molecular Probes (Invitrogen, Carlsbad, CA, USA).

All chemicals were purchased from Sigma–Aldrich (St Louis, MO, USA) unless otherwise noted. Complete mini EDTA-free, protease inhibitor cocktail tablets were from Roche diagnostics (Mannheim, Germany). Tris/glycine buffer (10X), Tris/glycine/SDS buffer (10X), and Precision Plus Protein All Blue Standards (Catalog #161–0373) were obtained from Bio-Rad (Hercules, CA) and anti-AMPKα from Millipore (Billerica, MA), anti-phospho-Thr172-AMPKα, anti-CaMKIV, and anti-rabbit IgG (H+L) (DyLight 680 Conjugate) from Cell Signaling Technology, Inc (Danvers, MA). Anti- CaMKKβ and anti-CaMKKα were obtained from Abgent INC (San Diego, CA). Anti-CaMKI and anti-phospho-Thr177-CaMKI were obtained from Novus Europe (Cambridge, UK). Anti-phospho-Thr196-CaMKIV was obtained from Santa Cruz Biotechnology, INC (Texas, USA). PI/Sybr-14 (LIVE/DEAD sperm viability kit) was purchased from Molecular Probes (Saint Aubin, France).